Dogwood In The Three Kinds Of Iridoid Terpene Glycosides Microwave Extraction And Hplc Determination

Cornus officinalis is the ripe fruit of cornet that belongs to cornet genus, cornel family. It is one of the 40 key crude medicinal materials of large consumption. The earliest record about cornel is in shen nong ben cao jing, a legendary medical book in China. The medicinal cornel is distributed in the north temperate zone and subtropical alpine area naturally. It is distributed widely in Zhejiang, Henan, Anhui, Shaanxi, Sichuan of our country. The Cornus officinalis, which is the astringent analeptics, has the effects of solid prostration, the astringent essence, and the hepatorenal with the taste of bitter and nature micro warm.The total saponin, in which there are about ten iridoid glycosides including loganin, sweroside, cornuside and so on, is one kind of main active components of corpus. Large researches show that saponin has the biological effect of dispelling sputum, relieving a cough, restraining bacterium, resisting cancer, reducing fever and so on. So it has broad development prospect. The loganin has the bidirectional control action to the immune response, and have certain protective function to the galactosamine result liver damage, the ascites lump cell, the lymphocyte, human bones medullary sarcoma cell SP20. Sweroside has the anti-liver cell damage function. The Chinese pharmacopoeia of 2005 version announces that the loganin's content in Cornus officinalis is one of Cornus officinalis raw material for medicine quality control targets. At present, many literatures appraisal Cornus officinalis raw material for medicine quality just with loganin content. Li Jun-Song et al determined the loganin and sweroside content in the Cornus officinalis used the highly effective capillary vessel electrophoresis. Author think it will be more all-round and reasonable to simultaneously determine the content of loganin, sweroside and cornuside in Cornus officinalis raw material for medicine quality. In this paper, the microwave-assisted extraction conditions of loganin, sweroside and cornuside in Cornus officinalis was optimized by means of response surface methodology (RSM), HPLC separation conditions of the iridoid glycosides were researched and their contents were determined by HPLC. The consequences are as follows:1. The optimal conditions of microwave extraction are as follows:microwave power 400 W, ethanol concentration 72%, liquid to sample ratio 15 mL/g, the extraction time 10 min, the extraction times 2.2. Three constituents were separated on an Agilent TC-C 18(250 mm×4.6 mm i.d,) column by gradient elution using acetonitrile and water as the mobile phase. The flow rate was 1.0 ml/min. The detection wave length was set at 240nm. The column temperature was set at 30℃.3. Loganin, sweroside and cornuside's retention time respectively are in turn 12.528 min,13.036 min and 17.252 min under the elected chromatograph condition. All the constituents showed good linearity (r>0.9994) in the range of the tested concentration, the average recoveries of the method were 99.35%,98.40% and 98.79% respectively.There are four parts in this thesis.Part one:reviewFirstly, it reviewed the research survey, chemical composition, pharmacology function, research present situation and significance and so on of Cornus officinalis; Secondly, it summaried the chemical composition, the methods of extraction and purification, the methods of content determination of Cornus officinalis total soap glucoside; Lastly, it briefly summaried the development research of extraction methods for traditional Chinese medicine.Part two:the extraction technology of the three iridoid glycosidesThe 5.0 g Cornus officinalis sample powder which had siftered through 40 meshes was weighed and put in the round flask, then added 72% ethyl alcohol 50mL into the round flask microwaving (400 W), withdrawed in 10 min, Taked out the upper formation solution, added 72% ethyl alcohol 50 mL to the residual again, repeated the operates one times as before, merged the filtrate, cleaned the filter paper and the residual with 72% ethyl alcohol, merged the cleaning liquid into the fluid, after revolving evaporation and freeze-drying, the crude extract was obtained, dissolved it to certain volume and determined the content for extraction yield. The microwave-assisted extraction conditions of loganin, sweroside and cornuside in Cornus officinalis was optimized by means of response surface methodology (RSM).Part three:content determination of the three iridoid glycosides by HPLCThe three iridoid glycosides in cornus were extracted in the optimal conditions and Their content were determined by HPLC.Part four:research summaryIn this part, it summaried the research content and predicted its future development.